Biofilm disruptive compositions

ABSTRACT

In certain embodiments, the present invention relates to compounds, compositions, and methods for disrupting biofilms. In some embodiments, the compounds and compositions comprise unsaturated long chain alcohols and/or aldehydes, or combinations of such compounds. In further embodiments, the present invention contains therapeutic actives to help reduce and/or eradicate the bacteria in the biofilm once the film is disrupted.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. non provisional applicationSer. No. 12/623,881 filed Nov. 23, 2009, the complete disclosure ofwhich is hereby incorporated herein by reference for all purposes.

FIELD OF INVENTION

In certain embodiments, the present invention relates to compounds,compositions, and methods for disrupting biofilms. In particularembodiments, the present invention relates to the use of unsaturatedlong chain alcohols and/or aldehydes, or combinations of such compoundsin compositions for disrupting biofilms.

BACKGROUND OF THE INVENTION

Biofilms are mucilaginous communities of microorganisms such asbacteria, archaea, fungi, molds, algae or protozoa or mixtures thereofthat grow on various surfaces. Biofilms form when microorganismsestablish themselves on a surface and activate genes involved inproducing a matrix that includes polysaccharides. This matrix mayprovide protection of biofilm forming bacteria from biocides.

Under certain circumstances, however, biofilms can be undesirable. Forexample, biofilms can cause damage to equipment such as cooling systems,or aquaculture equipment by corrosion of the equipment by microorganismsresiding in the biofilm or by excessive coating or film buildupcompromising the normal mechanics of the equipment. Biofilms can alsohave very detrimental health effects. For example, manyhospital-acquired infections involve biofilms, which can contaminateimplants and catheters and prevent adequate antimicrobial treatment ofsuch devices. Biofilms also result in adverse health conditions rangingfrom lung infections as in cystic fibrosis to more prevalent diseasessuch as tooth decay.

Regarding oral biofilms, certain bacteria can produce highly branchedpolysaccharides, which together with other microorganisms from the oralcavity form adhesive matrix films facilitating the proliferation ofplaque. Left untreated, these oral biofilms can eventually lead todental caries, gingival inflammation, periodontal disease, and toothloss. As oral biofilm continues to accumulate, rock-hard white oryellowish deposits can arise. These deposits are called calcifiedplaque, calculus or tartar, and are formed in the saliva from plaque andminerals, in particular calcium. Established oral biofilm can be verydifficult to disrupt whether by mechanical or chemical means. This canbe particularly problematic in domains where mechanical removal is notfeasible (depending on the implement).

Mechanical removal is an effective methodology used to dispersebiofilms. In the case of oral biofilms, toothbrushes, floss, picks, etc.have been used. One limitation of mechanical removal is the ability ofthe mechanical action to reach the biofilm coated surfaces. This isparticularly difficult between teeth and at the tooth/gum lineinterface.

Another method for preventing or disrupting a biofilm is to interferewith the quorum-sensing signals. Quorum-sensing signals are moleculesthat help trigger and coordinate part of the process of forming abiofilm. Bacteria constantly secrete low levels of the signals and sensethem either through receptors on their surfaces, or internally. Thereceptors trigger behavioral changes when there are enough bacteria toallow the signals' concentrations to achieve a critical threshold. Oncethis occurs, bacteria respond by adopting communal behavior, such asforming a biofilm, and in the case of pathogenic bacteria, deployingvirulence factors such as toxins. In addition to communicating withmembers of their own species, bacteria also conduct interspeciescommunications, such that a biofilm may involve and/or contain more thanone species of bacteria. Chemicals have been developed that bind butfail to activate the receptors of quorum-sensing signals or thatinterfere with signal synthesis. Enzymes that degrade the signals havealso been developed.

Strong antimicrobials may be used to kill bacteria in a biofilm,controlling its development and growth. However, once biofilms areestablished, antimicrobials are not associated with removal of live ordead biofilm. It has been well documented that, because antimicrobialshave difficulty penetrating the biofilm's surface layer, they are lesseffective on bacteria in an established biofilm compared to planktonicbacteria. Agents that help antimicrobials penetrate the biofilm'ssurface layer improve the effectiveness of the antimicrobials.

Recently, several biofilm dispersing agents have been identified thatcan be used to signal select bacteria to release from an establishedbiofilm. Signaling agents could help to disperse bacteria, however,signaling compounds have not been shown to be effective on all oralbacterial species, and often need optimized conditions and long contacttimes to show functionality.

There is, therefore, an ongoing need to identify agents that function ascompounds, compositions, and methods for disrupting biofilms. In certainembodiments, the present invention provides certain unsaturated longchain alcohols and/or aldehydes which disrupt biofilms and dispersesingle and mixed species bacteria from the biofilm.

SUMMARY OF THE INVENTION

In certain embodiments, the present invention is related tocompositions, and methods for disrupting biofilms.

In particular embodiments, the present invention comprises a biofilmdisruptor comprising at least one unsaturated aliphatic long chainalcohols and/or aldehydes, or combinations of such compounds. In stillother embodiments, the present invention may further contain therapeuticactives to facilitate the reduction and/or eradication of bacteria inthe biofilm once the film is disrupted.

In one embodiment, the present invention relates to the use as a biofilmdisruptor of at least one unsaturated, aliphatic long chain alcoholand/or aldehyde of formula:R¹(C_(n)R² _(s)H_((2n-2m-s)))R³(C_(x)R⁴ _(t)H_((2x-2y-t)))R⁵CH₂R⁶wherein R¹ and R⁶ are, independently, H—, HO—, O═CH—, CH₃— or CH₂═CH—,(—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷, (—OCH₂CH(CH₃)—)_(z)R⁷; R² and R⁴ are,independent of themselves and independent of each other, HO—, O═CH—,branched or straight chain C₁ to C₄ alkyl or alkene, —CH₂OH, —CH₂CH₂OH;R³ is, independently, single bond or —O—; R⁵ is, independently, singlebond, —O—, (—OCH₂—)_(z), (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z); R⁷ is,independent of itself, H—, HO—, O═CH—, CH₃— or CH₂═CH—; n is an integerfrom 1 to 10; m (the degree of unsaturation) is an integer from 0 to n/2if n is even or (n−1)/2 if n is odd; x is an integer from 0 to 10; y(the degree of unsaturation) is an integer from 0 to x/2 if x is even or(x−1)/2 if x is odd and z is an integer from 0 to 5; s is an integer≦n;and t is an integer≦x, provided that:

-   -   if neither R¹, R², R⁴, nor R⁶ contain any C═C unsaturation, then        m+y is at least 1;    -   R⁵ and R⁶ are not such that R⁵ is (—OCH₂—)_(z), (—OCH₂CH₂—)_(z),        (—OCH₂CH(CH₃)—)_(z), and R⁶ is (—OCH₂—)_(z)R⁷,        (—OCH₂CH₂—)_(z)R⁷, (—OCH₂CH(CH₃)—)_(z)R⁷;    -   The carbon-carbon chain length from R¹ to R⁶ ranges continuously        or intermittently from at least 7, optionally from 8 to 13        carbons, or optionally, from 9 to 12; and    -   at least one of any one of R¹, R², R⁴ and R⁶ contains HO— or        O═CH—.

In a second embodiment, the present invention relates to a compositionfor disrupting biofilms, comprising:

-   -   i. from about 0.005% to about 10% of a biofilm disruptor        comprising at least one unsaturated aliphatic long chain alcohol        and/or aldehyde of the formula:        R¹(C_(n)R² _(s)H_((2n-2m-s)))R³(C_(x)R⁴        _(t)H_((2x-2y-t)))R⁵CH₂R⁶        -   wherein R¹ and R⁶ are, independently, H—, HO—, O═CH—, CH₃—            or CH₂═CH—, (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,            (—OCH₂CH(CH₃)—)_(z)R⁷; R² and R⁴ are, independent of            themselves and independent of each other, HO—, O═CH—,            branched or straight chain C₁ to C₄ alkyl or alkene, —CH₂OH,            —CH₂CH₂OH; R³ is, independently, single bond or —O—; R⁵ is,            independently, single bond, —O—, (—OCH₂—)_(z),            (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z); R⁷ is, independent of            itself, H—, HO—, O═CH—, CH₃— or CH₂═CH—; n is an integer            from 1 to 10; m (the degree of unsaturation) is an integer            from 0 to n/2 if n is even or (n−1)/2 if n is odd; x is an            integer from 0 to 10; y (the degree of unsaturation) is an            integer from 0 to x/2 if x is even or (x−1)/2 if x is odd            and z is an integer from 0 to 5; s is an integer≦n; and t is            an integer≦x, provided that:            -   if neither R¹, R², R⁴, nor R⁶ contain any C═C                unsaturation, then m+y is at least 1;            -   R⁵ and R⁶ are not such that R⁵ is (—OCH₂—)_(z),                (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z), and R⁶ is                (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,                (—OCH₂CH(CH₃)—)_(z)R⁷;            -   the carbon-carbon chain length from R¹ to R⁶ ranges                continuously or intermittently from at least 7,                optionally from 8 to 13 carbons, or optionally, from 9                to 12; and            -   at least one of any one of R¹, R², R⁴ and R⁶ contains                HO— or O═CH—;    -   ii. optionally, a therapeutic active; and    -   iii. a carrier selected from the group consisting of a non-oral        carrier, pharmaceutically acceptable carrier, orally acceptable        carrier and dermatogically acceptable carrier.

In a further embodiment, the present invention relates to an article ordevice, comprising:

-   -   i. an article or device;    -   ii. from about 0.005% to about 10% of a biofilm disruptor        applied to the device or article, the biofilm disruptor        comprising at least one unsaturated aliphatic long chain alcohol        and/or aldehyde of the formula:        R¹(C_(n)R² _(s)H_((2n-2m-s)))R³(C_(x)R⁴        _(t)H_((2x-2y-t)))R⁵CH₂R⁶        -   wherein R¹ and R⁶ are, independently, H—, HO—, O═CH—, CH₃—            or CH₂═CH—, (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,            (—OCH₂CH(CH₃)—)_(z)R⁷; R² and R⁴ are, independent of            themselves and independent of each other, HO—, O═CH—,            branched or straight chain C₁ to C₄ alkyl or alkene, —CH₂OH,            —CH₂CH₂OH; R³ is, independently, single bond or —O—; R⁵ is,            independently, single bond, —O—, (—OCH₂—)_(z),            (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z); R⁷ is, independent of            itself, H—, HO—, O═CH—, CH₃— or CH₂═CH—; n is an integer            from 1 to 10; m (the degree of unsaturation) is an integer            from 0 to n/2 if n is even or (n−1)/2 if n is odd; x is an            integer from 0 to 10; y (the degree of unsaturation) is an            integer from 0 to x/2 if x is even or (x−1)/2 if x is odd            and z is an integer from 0 to 5; s is an integer≦n; and t is            an integer≦x, provided that:            -   if neither R¹, R², R⁴, nor R⁶ contain any C═C                unsaturation, then m+y is at least 1;            -   R⁵ and R⁶ are not such that R⁵ is (—OCH₂—)_(z),                (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z), and R⁶ is                (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,                (—OCH₂CH(CH₃)—)_(z)R⁷;            -   the carbon-carbon chain length from R¹ to R⁶ ranges                continuously or intermittently from at least 7,                optionally from 8 to 13 carbons, or optionally, from 9                to 12; and            -   at least one of any one of R¹, R², R⁴ and R⁶ contains                HO— or O═CH—; and    -   iii. optionally, a therapeutic active applied to the device or        article.

In yet another embodiment, the present invention relates to a method ofdisrupting biofilms, comprising the steps of:

-   -   i. providing a surface or substrate comprising a biofilm; and    -   ii. applying to the surface or substrate from about 0.005% to        about 10% of a biofilm disruptor comprising at least one        unsaturated aliphatic long chain alcohol and/or aldehyde of the        formula:        R¹(C_(n)R² _(s)H_((2n-2m-s)))R³(C_(x)R⁴        _(t)H_((2x-2y-t)))R⁵CH₂R⁶        -   wherein R¹ and R⁶ are, independently, H—, HO—, O═CH—, CH₃—            or CH₂═CH—, (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,            (—OCH₂CH(CH₃)—)_(z)R⁷; R² and R⁴ are, independent of            themselves and independent of each other, HO—, O═CH—,            branched or straight chain C₁ to C₄ alkyl or alkene, —CH₂OH,            —CH₂CH₂OH; R³ is, independently, single bond or —O—; R⁵ is,            independently, single bond, —O—, (—OCH₂—)_(z),            (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—); R⁷ is, independent of            itself, H—, HO—, O═CH—, CH₃— or CH₂═CH—; n is an integer            from 1 to 10; m (the degree of unsaturation) is an integer            from 0 to n/2 if n is even or (n−1)/2 if n is odd; x is an            integer from 0 to 10; y (the degree of unsaturation) is an            integer from 0 to x/2 if x is even or (x−1)/2 if x is odd            and z is an integer from 0 to 5; s is an integer≦n; and t is            an integer≦x, provided that:            -   if neither R¹, R², R⁴, nor R⁶ contain any C═C                unsaturation, then m+y is at least 1;            -   R⁵ and R⁶ are not such that R⁵ is (—OCH₂—)_(z),                (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z), and R⁶ is                (—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷,                (—OCH²CH(CH³)—)_(z)R⁷;            -   the carbon-carbon chain length from R¹ to R⁶ ranges                continuously or intermittently from at least 7,                optionally from 8 to 13 carbons, or optionally, from 9                to 12; and            -   at least one of any one of R¹, R², R⁴ and R⁶ contains                HO— or O═CH—.

In certain embodiments, this invention is also directed to methods fordisrupting biofilms on the surface of living entities and/or non-livingthings. In other embodiments, the alcohols and/or aldehydes disclosedherein are contacted with the surface of the biofilm in atherapeutically effective amount to disrupt the biofilm, and facilitatethe reduction and/or eradication of the bacteria in the biofilm once thefilm is disrupted.

DETAILED DESCRIPTION OF THE INVENTION

The compositions of the present invention can comprise, consist of, orconsist essentially of the essential elements and limitations of theinvention described herein, as well any of the additional or optionalingredients, components, or limitations described herein.

The term “comprising” (and its grammatical variations) as used herein isused in the inclusive sense of “having” or “including” and not in theexclusive sense of “consisting only of.” The terms “a” and “the” as usedherein are understood to encompass the plural as well as the singular.

All documents cited are, in relevant part, incorporated herein byreference; the citation of any document is not to be construed as anadmission that it is prior art with response to the present invention.

The present invention provides compounds, compositions, and methods fordisrupting biofilms on the surface of living entities and/or non-livingthings. The invention also provides methods for removing oral biofilmsfrom the surface of living entities and/or non-living things, andpreventing the re-establishment of biofilms by facilitating thereduction and/or eradication of any bacteria before the film can bereestablished.

The term “biofilm” means a mucilaginous community of microorganisms suchas bacteria, archaea, fungi, molds, algae or protozoa or mixturesthereof that grow on various surfaces when the microorganisms establishthemselves on a surface and activate genes involved in producing amatrix that includes polysaccharides.

The phrase “oral biofilm” means a mixture of bacteria, epithelial cells,leukocytes, macrophages and other oral exudates that forms on thesurface of unclean teeth. An oral biofilm can comprise one or more thanone species of bacteria.

The phrase “therapeutically effective amount” means the concentration orquantity or level of the compound of the present invention that canattain a particular medical end in disrupting biofilms, such asdecreasing biofilm formation, dispersing biofilms, or having toxicactivity for biofilms.

The phrase “non-oral and non-dermatological carrier” means that thecarrier is not suitable for oral ingestion or application to thesurfaces of the oral cavity, skin and/or mucosal surfaces of livingorganisms including, but not limited to mammals or humans application oringestion by a mammal without undue toxicity, incompatibility,instability, allergic response, and the like.

The phrase “pharmaceutically acceptable” means that the drugs,medications or inert ingredients which the term describes are suitablefor use in humans and lower animals without undue toxicity,incompatibility, instability, irritation, allergic response, and thelike.

The phrase “orally acceptable” means that the carrier is suitable forapplication to the surfaces of the oral cavity or ingestion by a livingorganism including, but not limited to, mammals and humans without unduetoxicity, incompatibility, instability, allergic response, and the like.

The phrase “dermatologically acceptable carrier” means that the carrieris suitable for topical application to the keratinous tissue, has goodaesthetic properties, is compatible with the actives of the presentinvention and any other components, and will not cause any safety ortoxicity concerns.

The terms “disrupter”, “disruptive” or “disruption” means the partial orcomplete removal of biofilm or biofilm matrix and/or compromise theintegrity of the biofilm.

Biofilm Disruptor

In certain embodiments, the present invention incorporates and/orrelates to the use of a biofilm disruptor. The biofilm disruptors of thepresent invention are or comprise at least one unsaturated aliphaticlong chain alcohol or aldehyde of the formula:R¹(C_(n)R² _(s)H_((2n-2m-s)))R³(C_(x)R⁴ _(t)H_((2x-2y-t)))R⁵CH₂R⁶wherein R¹ and R⁶ are, independently, H—, HO—, O═CH—, CH₃— or CH₂═CH—,(—OCH₂—)_(z)R⁷, (—OCH₂CH₂—)_(z)R⁷, (—OCH₂CH(CH₃)—)_(z)R⁷; R² and R⁴ are,independent of themselves and independent of each other, HO—, O═CH—,branched or straight chain C₁ to C₄ alkyl or alkene, —CH₂OH, —CH₂CH₂OH;R³ is, independently, single bond or —O—; R⁵ is, independently, singlebond, —O—, (—OCH₂—)_(z), (—OCH₂CH₂—)_(z), (—OCH₂CH(CH₃)—)_(z); R⁷ is,independent of itself, H—, HO—, O═CH—, CH₃— or CH₂═CH—; n is an integerfrom 1 to 10; m (the degree of unsaturation) is an integer from 0 to n/2if n is even or (n−1)/2 if n is odd; x is an integer from 0 to 10; y(the degree of unsaturation) is an integer from 0 to x/2 if x is even or(x−1)/2 if x is odd and z is an integer from 0 to 5; s is an integer≦n;and t is an integer≦x, provided that:

-   -   if neither R¹, R², R⁴, nor R⁶ contain any C═C unsaturation, then        m+y is at least 1;    -   R⁵ and R⁶ are not such that R⁵ is (—OCH₂—)_(z), (—OCH₂CH₂—)_(z),        (—OCH₂CH(CH₃)—)_(z), and R⁶ is (—OCH₂—)_(z)R⁷,        (—OCH₂CH₂—)_(z)R⁷, (—OCH₂CH(CH₃)—)_(z)R⁷;    -   the carbon-carbon chain length from R¹ to R⁶ ranges continuously        or intermittently from at least 7, optionally from 8 to 13        carbons, or optionally, from 9 to 12; and    -   at least one of any one of R¹, R², R⁴ and R⁶ contains HO— or        O═CH—.

In certain embodiments, R¹, R², R⁴ and R⁶ are not such that one of R¹,R², R⁴ and R⁶ is O═CH— and any other one of R¹, R², R⁴ and R⁶ is HO—.

In some embodiments, at least two of any one of, or optionally, at leastone of any one of R¹, R², R⁴ and R⁶ is HO— or O═CH—. In otherembodiments, R³ and R⁵ are single bonds. In still other embodiments, atleast one of any one of R¹, R², R⁴ and R⁶ is HO—. In an alternativeembodiment, at least one of any one of R¹, R², R⁴ and R⁶ is O═CH—.

In certain embodiments, s+t is at least 1. In certain other embodiments,the biofilm disruptor is a linear unsaturated aliphatic long chainalcohol or aldehyde.

In certain embodiments, the biofilm disruptor comprises a compoundselected from the group consisting of unsaturated long chain alcoholsand/or aldehydes of formulas: CH₃(C_(n)R¹ _(s)H_((2n-2m-s)))CH₂R²,CH₂—CH(C_(n)R¹ _(s)H_((2n-2m-s)))CH₂R² and mixtures thereof wherein eachR¹ and R² are, independent of themselves and independent of each other,H—, HO—, O═CH—, branched or straight chain C₁ to C₄ alkyl or alkene,—CH₂OH, —CH₂CH₂OH; and n is an integer from 5 to 11; m is an integerfrom 1 to 3 and s is an integer≦n, provided that at least one of any oneof R¹ and R² contains HO— or O═CH— and provided that R¹ and R² are notsuch that one contains O═CH— and the other contains HO—.

In still other embodiments, the biofilm disruptor comprises a compoundselected from the group consisting of unsaturated long chain alcoholsand/or aldehydes of formulas: CH₃(C_(n)R¹ _(s)H_((2n-2m-s)))CH₂R²,CH₂═CH(C_(n)R¹ _(s)H_((2n-2m-s)))CH₂R² and mixtures thereof wherein eachR¹ and R² are, independent of themselves and independent of each other,H—, HO—, O═CH—, branched or straight chain C₁ to C₄ alkyl or alkene,—CH₂OH, —CH₂CH₂OH; and n is an integer from 5 to 11; and s is aninteger≦n, provided that at least one of any one of R¹ and R² containsHO— or O═CH— and provided that R¹ and R² are not such that one containsO═CH— and the other contains HO—.

In certain other embodiments, the biofilm disruptor is an unsaturatedalcohol or aldehyde selected from the group consisting of 1-decen-3-ol;cis-4-decen-1-ol, trans-2-decen-1-ol, cis-2-nonen-1-ol, cis-4-decenal,trans-2-decenal, cis-7-decenal, cis-5-octen-1-ol, trans-2-octen-1-ol,1-octen-3-ol, cis-3-nonen-1-ol, trans-2-nonen-1-ol, cis-6-nonen-1-ol,9-decen-1-ol, trans-2-undecen-1-ol, trans-2-dodecen-1-ol,trans-2-octenal, trans-2-nonenal, 6-nonenal, cis-2-decenal,trans-2-undecenal, trans-2-dodecenal, cis-3-octen-1-ol, 3-octen-2-ol,10-undecen-1-ol, trans-2-tridecen-1-ol, stereoisomers thereof andmixtures thereof.

In other embodiments, the biofilm disruptor is a unsaturated alcoholselected from the group consisting of cis-2-nonen-1-ol, 1-decen-3-ol,stereoisomers thereof and mixtures thereof. In still other embodiments,the biofilm disruptor is 1-decen-3-ol.

In alternative embodiments, the biofilm disruptor is a unsaturatedaldehyde selected from the group consisting of trans-2-dodecenal,cis-4-decenal, trans-2-decenal, trans-2-nonenal, trans-2-undecenal,stereoisomers thereof and mixtures thereof. In some embodiments, thebiofilm disruptor is an unsaturated aldehyde selected from the groupconsisting of trans-2-dodecenal, cis-4-decenal, trans-2-decenal,trans-2-nonenal, stereoisomers thereof and mixtures thereof. In otherembodiments, the biofilm disruptor is an unsaturated aldehyde selectedfrom the group consisting of trans-2-dodecenal, cis-4-decenal,trans-2-decenal, trans-2-nonenal, stereoisomers thereof and mixturesthereof. In still other embodiments, the biofilm disruptor is aunsaturated aldehyde selected from the group consisting oftrans-2-dodecenal, cis-4-decenal, trans-2-decenal, stereoisomers thereofand mixtures thereof.

In certain other embodiments, the present invention also providesmethods for disrupting biofilms. In such nonlimiting embodiments, themethod comprises contacting a surface or substrate with atherapeutically effective amount of the biofilm disruptors orcompositions described above. In certain embodiments of the invention,the surface or substrate is treated with the biofilm disruptors of thepresent invention by applying the biofilm disruptor onto a surface orsubstrate so as to leave a residue or deposit of the biofilm disruptoron the surface or substrate that remains of the surface or substrate forat least about 1 (or about 1), optionally 4 (or about 4), optionally 10(or about 10), optionally 30 (or about 30), optionally 60 (or about 60)or optionally 120 (or about 120) seconds. In some embodiments, thebiofilm disruptor is impregnated in a surface in order to inhibitformation of a biofilm on the surface. In alternative embodiments, thebiofilm disruptor can be in a copolymer or a gel coating over thesurface.

In the case of certain oral care or orally ingestible compositionembodiments, the biofilm disruptor of the present invention is presentat a level of from 0.005% (or about 0.005%) to 0.5% (or about 0.5%),optionally from 0.01% (or about 0.01%) to 0.3% (or about 0.3%),optionally from 0.025% (or about 0.025%) to 0.2% (or about 0.2%), oroptionally from 0.05% (or about 0.05%) to less than 0.1% (or about0.1%).

In the case of certain composition embodiments for dermatological useand/or for coating onto or disrupting biofilms on non-oral andnon-dermatological surfaces, the biofilm disruptor of the presentinvention is present at a level of from 0.005% (or about 0.005%) to 10%(or about 10%), optionally from 0.01% (or about 0.01%) to 8% (or about8%), optionally from 0.025% (or about 0.025%) to 6% (or about 6%), oroptionally from 0.05% (or about 0.05%) to less than 4% (or about 4%).

Therapeutic Actives

In certain embodiments, the present invention further incorporates oruses in conjunction with the biofilm disruptor an additional therapeuticactive. In some embodiments, the active includes, but is not limited to,diffusible signals, antimicrobial actives, anti-inflammatory actives,oral care actives, skin care actives and mixtures thereof.

Diffusible Signals

Diffusible signals are molecules synthesized, released and/or detectedby microbial cells as a mechanism for cell-cell communication.Diffusible signals useful in the compositions of the present inventioninclude, but are not limited to, at least one N-acylhomoserine lactonesof formula

where R is

at least one autoinducer oligopeptides such as

ADITROWGD (ComX [B. subtilis]), ERGMT (CSF [B. subtilis]),EMRLSKFFRDFILQRKK (CSP [S. pneumonia]); at least one autoinducer-2compounds such as

at least one Streptomyces butryolactores such as

at least one

at least one diketopiperzines such as

at least one diffusible signal factor (DSF) such as, but are not limitedto, cis-2-dodecenoic acid (BDSF B. cenocepacia]),cis-11-methyl-2-dodecenoic acid (DSF [X. campestris]),12-methyltetradecanoic acid (Xf DSF [X fastidiosa]); at least one DSFderivative including, but are not limited to, cis or trans 2-decenoicacid, cis or trans 2-hexenoic acid, cis or trans 2-heptenoic acid, cisor trans 2-octenoic acid, cis or trans 2-nonenoic acid, cis or trans2-undecenoic acid, cis or trans 2-dodecenoic acid, cis or trans2-tridecenoic acid, cis or trans 2-tetradecenoic acid, cis or trans2-pentadecenoic acid, cis or trans 2-hexadecenoic acid, cis or trans2-heptadecenoic acid, cis or trans 2-octadecenoic acid, cis or trans2-nonadecenoic acid. Organisms recited above in parentheses describe theorganisms that synthesize, release and/or detect the specified signalmolecule. A more detailed discussion of DSF or DSF-like compounds can befound in US patent publications US20060260007 to Wang et al. andUS20080317815 to Davies, both of which are herein incorporated byreference in their entirety. A more detailed discussion of diffusiblesignal factors generally can be found in R P Ryan and J M Dow (2008),“Diffusable signals and interspecies communication in bacteria”,Microbiology, 154, 1845-1858.Antimicrobial Actives

Antimicrobial active suitable for use herein include, but are notlimited to, triclosan, metronidazole, tetracyclines, quinolones, plantessential oils, camphor, thymol, carvacrol, menthol, eucalyptol, methylsalicylate, tobramycin, cetylpyridinium chloride, neomycin, polymyxin,bacitracin, clindamycin, ciprofloxacin, rifampin, oxfloxacin,macrolides, penicillins, cephalosporins, amoxicillin/clavulanate,quinupristin/dalfopristin, amoxicillin/sulbactum, fluoroquinolones,ketolides, aminoglycosides and mixtures thereof.

Anti-Inflammatory Actives

In certain embodiments, the compositions of the present inventionfurther contain anti-inflammatory actives. Anti-inflammatory activesuseful in the present invention include steroidal anti-inflammatoryactives, non-steroidal anti-inflammatory actives and mixtures thereof.Suitable steroidal anti-inflammatory include, but not limited to,hydrocortisone, fluocinolone acetonide, halcinonide, halobetasolpropionate, clobetasol propionate, betamethasone dipropionate,betamethasone valerate, triamcinolone acetonide and mixtures thereof.Suitable non-steroidal anti-inflammatory actives include, but are notlimited to, salicylic acid derivatives such as aspirin, sodiumsalicylate, choline magnesium salicylate, salsalate, diflunisal,salicylsalicylic acid, sulfasalazine, and olsalazine; para-aminophenolderivatives such as acetaminophen; indole and indene acetic acids suchas indomethacin, sulindac, and etodolac; heteroaryl acetic acids such astolmetin, diclofenac, and ketorolac; arylpropionic acids such asibuprofen, naproxen, flurbiprofen, ketoprofen, fenoprofen, andoxaprozin; anthranilic acids (fenamates) such as oxicams (piroxicam,tenoxicam), pyrazolidineones (phenylbutazone, oxyphenthatrazone);alkanones such as nabumetone; apazone (azapropazone); nimesulide; andmixtures thereof.

Oral Care Actives

In certain embodiments, the compositions of the present inventionfurther contain oral care actives. In certain embodiments, the oral careactives include, but are not limited to, essential oils (such asmenthol, methyl salicylate, eucalyptol, thymol and mixtures thereof),anti-plaque agents, fluoride ion sources such as sodium fluoride, sodiummonofluorophosphate, stannous fluoride, and amine fluorides (providing,for example, about 1-5000 ppm of fluoride ion, optionally about 200-1150ppm of fluoride ion); anti-calculus agents such as water-solublepyrophosphate salts, preferably alkali metal pyrophosphates; chelatingagents; tooth desensitization agents which reduce tooth sensitivityincluding potassium salts such as potassium nitrate and potassiumchloride (for example about 1% to about 5% by weight) and strontiumsalts such as strontium chloride and strontium acetate (for exampleabout 2% to about 10% by weight); tooth whitening agents and vitaminssuch as vitamin A.

In certain embodiments, suitable anti-plaque agents include, but are notlimited to, non-ionic antibacterial agents such as bromochlorophene andtriclosan and cationic agents such as cetylpyridinium chloride andchlorhexidine salts, and mixtures thereof. In general, non-ionicantibacterial agents have a very low solubility in water and have notbeen incorporated into mouthwash preparations other than thosecontaining high levels of alcohol. Furthermore, it is known that certainwater-insoluble flavoring oils such as anethole and menthol have anantibacterial effect at high concentrations. In other embodiments, amajor advantage provided by oral care compositions of the presentinvention is that it allows for the incorporation into non-alcoholiccompositions of water-insoluble antibacterial agents and/orwater-insoluble anti-calculus agents at effective levels as emulsions orsuspensions. In certain embodiments, the oral care compositions of thepresent invention comprise from about 0.001% to about 1%, optionallyfrom about 0.01% to about 0.5% by weight of a non-ionic antibacterialagent. In some embodiments, the water-insoluble anti-tartar agentscomprise zinc salts such as zinc citrate. In certain embodiments, thecompositions of the present invention can comprise from about 0.1% toabout 1% of a water-insoluble anti-calculus agent.

A more detailed discussion of abrasives, surfactants oral care activesand/or other ingredients useful in the compositions of the presentinvention can be found in U.S. Pat. No. 7,025,950 to Majeti et al.: U.S.Pat. No. 6,682,722 to Majeti et al.; U.S. Pat. No. 6,782,307 to Wilmottet al. and U.S. Pat. No. 6,121,315 to Nair et al., each of which isherein incorporated by reference in its entirety.

Skin Care Actives

In certain embodiments, the compositions of the present inventionfurther contain skin care actives. In certain embodiments, the oral careactives include, but are not limited to alpha- or beta-hydroxy acids,and derivatives, salts, isomers and tautomers thereof. Non-limitingexamples of alpha- and beta-hydroxy acids include salicylic acid,alpha-hydroxy-butyric acid, alpha-hydroxyisobutyric acid,alpha-hydroxyisocaproic acid, alpha-hydroxyisovaleric, atrolactic acid,beta-hydroxybutyric acid, beta-phenyl lactic acid, beta-phenylpyruvicacid, citric acid ethyl pyruvate, galacturonic acid, glucoheptonic acid,glucoheptono 1,4-lactone, gluconic acid, gluconolactone glucuronic acid,glucuronolactone, glycolic acid, isopropyl pyruvate, lactic acid, malicacid, mandelic acid, methyl pyruvate, mucic acid, pyruvic acid,saccharic acid, saccharic acid 1,4-lactone, tartaric acid and tartronicacid, and mixtures thereof.

Carriers

In certain embodiments, the fatty alcohols of the present invention arefurther mixed with appropriate carrier materials or ingredients to formdesired dosage forms. In some embodiments, the fatty alcohols andcompositions described herein may prepared as unitary dosage formssuitable for administration orally, percutaneously, by parenteralinjection (including subcutaneous, intramuscular, intravenous andintradermal), topically, intranasally, by inhalation, or for applicationto a medical device, such as an implant, catheter, or other device.

For pharmaceutically acceptable carriers that permit parenteraladministration, the pharmaceutically acceptable carriers often comprisesterile water, which may be supplemented with various solutes to, forexample, increase solubility. Injectable solutions may be prepared inwhich the pharmaceutically acceptable carrier comprises saline solution,glucose solution, or a mixture thereof, which may include certainwell-known anti-oxidants, buffers, bacteriostats, and other solutes thatrender the formulation isotonic with the blood of the intended patient.

For pharmaceutically acceptable carriers that permit intranasaladministration, the pharmaceutically acceptable carriers often comprisepoly acrylic acids such as Carbopol® 940, a hydrogenated castor oil suchas Cremophor® RH40, glycerol, vinylpyrrolidones such as PVP-K90® or PVPK30®, polyethylene glycols such as PEG 1450®, benzyl alcohol, Edetatesodium, hydroxycellulose, potassium chloride, potassium phosphate, andsodium phosphate. In some embodiments, the compositions used forintranasal administration also commonly include benzalkonium chloride asan anti-microbial preservative.

For pharmaceutically acceptable carriers that permit administration byinhalation, the pharmaceutically acceptable carriers often comprisesolvent/carrier/water mixtures that are easily dispersed and inhaled viaa nebulizer or inhaler. In some embodiments, for example, a mixture ofethanol/propylene glycol/water in the ratio of about 85:10:5 (partsethanol:parts propylene glycol:parts water) can be used to administerthe compounds and compositions of the invention via inhalation.

For orally acceptable carriers, the orally acceptable carrier may be inthe form of a mouthrinse, toothpaste, toothgel, dentifrice, breathspray, or prophylaxis paste, and comprise at least about 24%,optionally, at least about 60%, optionally, at least about 80% to about99%, or, optionally, at least about 80% to about 90% by weight of aliquid carrier. In certain embodiments, the liquid carrier may be in theform of a solution, emulsion or microemulsion of components and, in someembodiments, contain at least about 5% by weight water, optionally, atleast about 10% by weight water. In certain embodiments, alcohol such asethanol may optionally form part of the liquid carrier, for example,from about 5% to about 35% by weight of the liquid carrier, and, in someembodiments, is particularly useful in oral care compositions having ahigh flavor impact and breath-freshening and/or antiseptic properties.

In certain embodiments, the oral composition may also be in the form ofchewing gum, a breath strip, a lozenge, or a breath mint containing orcoated with a biofilm disruptor composition.

In other embodiments, the orally acceptable carriers are suspensions,syrups, elixirs and solutions. In alternate embodiments, the orallyacceptable carriers are solid carriers containing or coated with thelong chain alcohols, aldehydes or combinations thereof, including, butnot limited to, toothpowders, powders, whitening strip, breath strip,lozenge, pills, capsules, suppositories and tablets. In still otherembodiments, the solid carriers can be prepared so as to be suitable forrectal or vaginal application.

In certain embodiments, the pH of the oral care compositions accordingto the present invention is generally in the range of from about 3.5 toabout 9.0, optionally, from about 4.0 to 8.0, or optionally, from about4.0 to about 7.0. In other embodiments, if desired, the pH can becontrolled with acid, for example citric acid, or base, for examplesodium hydroxide, or buffered, for example with citrate, phosphate,benzoate or bicarbonate buffering salts.

In certain orally administered embodiments, sweeteners can be usedincluding, but not limited to, sucralose, aspartame, acesulfame K,saccharin, cyclamate and mixtures thereof.

Methods of Using the Biofilm Disrupters

In certain embodiments, the biofilm disruptors are used to treatarticles, devices, substrates and surfaces (mammalian or inanimate) todisrupt the formation of or disrupt already formed biofilms.

In some embodiments, the surface to be treated with the biofilmdisruptors includes medical devices such as catheters, respirators, andventilators. In other embodiments, the surface can be that of implantedmedical devices, including stents, artificial valves, joints, pins, boneimplants, sutures, staples, pacemakers, and other temporary or permanentmedical devices.

In other embodiments, the surface to be treated with the biofilmdisruptors includes articles such as drains, tubs, kitchen appliances,countertops, shower curtains, grout, toilets, industrial food andbeverage production facilities, flooring, and food processing equipmentand the like.

The surface to be treated with biofilm disruptors in yet anotherembodiment includes article surfaces such as filter or heat exchangersurfaces, providing means for reducing and/or eliminating biofouling ofheat exchangers or filters.

Other embodiments of the present invention, relate to use on orapplication of the biofilm disruptors to articles, devices, substratesor surfaces associated marine structures including, but not limited to,boats, piers, oil platforms, water intake ports, sieves, and viewingports.

In certain embodiments, the articles, substrate or device surface beingtreated with the biofilm disruptors can alternatively be associated witha system for water treatment and/or distribution (like drinking watertreatment and/or distributing systems, pool and spa water treatmentsystems, water treatment and/or distribution systems in manufacturingoperations, and a system for dental water treatment and/ordistribution). In some embodiments, the biofilm disruptor treatedarticle, substrate or device surface can also be associated with asystem for petroleum drilling, storage, separation, refining and/ordistribution (like petroleum separation trains, a petroleum container,petroleum distributing pipes, and petroleum drilling equipment). Inother embodiments, the biofilm disruptor can also be included informulations directed at reducing or eliminating biofilm deposits orbiofouling in porous medium, such as with oil and gas bearing geologicalformations. In particular embodiments, the biofilm disruptor treatmentmay be accomplished by applying a coating of the biofilm disruptor, suchas by painting, to the surface of articles, substrate or device.

In still other embodiments, the present invention further relates to amethod of using the biofilm disruptors to treat and/or prevent dentalplaque, dental carries, gingival disease, periodontal disease, and oralinfection in a subject. In such nonlimiting embodiments, the methodinvolves treating the surfaces of the oral cavity of the subject withthe biofilm disruptor according to the present invention. In particularembodiments, treatment can be carried out with a dentifrice, mouthwash,mouth rinse, dental floss, gum, strip, toothpaste, a toothbrushcontaining the biofilm disruptor, and other preparations containing thebiofilm disruptor. In certain other embodiments, the composition mayalso contain other compounds known in the dental arts that are typicallyadded to dental compositions. For example, in some embodiments, thebiofilm disruptor composition may also include such oral care actives asfluoride, desensitizing agents, anti-tartar agents, anti-bacterialagents, remineralization agents, whitening agents, abrasives andanti-caries agents.

In other embodiments, the biofilm disruptor is used with a dental deviceor article that is placed in the oral cavity. The biofilm disruptor iscoated on, encapsulated in, or impregnated in the dental article/device.Suitable dental articles/devices include, but are not limited to,dentures, dental dams, and certain types of orthodontic braces.Additional components or ingredients may be included in or with thebiofilm disruptor.

In certain other embodiments, the dental article is a dental floss. Anyfiber known in the art may be used in the dental floss. Suitable fibersinclude polyamides (such as nylon), polyesters, polypropylenes,polytetrafluoroethylenes, cellulose, and cotton. The biofilm disruptorcomposition may be impregnated into the fiber, coated on the fiber, orotherwise incorporated into the dental floss. In some embodiments, thedental floss may be coated or impregnated with a wax or otherhydrophobic substance for ease of use during the flossing process.Suitable waxes include microcrystalline waxes, beeswax, paraffin waxes,carnauba waxes, and polyethylene waxes. In certain embodiments, thebiofilm disruptor composition may be coated onto the dental floss aspart of the wax layer, as a second or additional layer in conjunctionwith the wax layer, or applied to the fiber as discussed above.

In certain embodiments, the dental article may be a toothpick that isimpregnated with or coated with the biofilm disruptor composition. Insome embodiments, the toothpicks may be made from natural products, suchas wood, or artificial components, including various plastics.

In certain embodiments, the dental article may also be a dentalappliance such as a dental aspirator, bite block, dental dam, tonguestabilizer, tongue deflector, or any other piece of dental equipmentthat a dentist or dental assistant may use in the mouth of a patient.The portion of the dental appliance that comes into contact with theoral cavity of a patient may be coated with the biofilm disruptorcomposition.

The dental article may also be a dental construct, such as a veneers,crowns, inlays, onlays, or bridges that are placed on the teeth. Dentalconstructs are typically made of metal alloys, porcelain, ceramic,amalgam, acrylate polymers, or a combination of these materials. Thebiofilm disruptor composition may be embedded in the composition used tomake the dental construct. Alternatively, the biofilm disruptorcomposition may be coated on the dental construct after it has beenprepared.

In some embodiments, the biofilm disruptor may be incorporated into thevarious parts of a toothbrush by means known in the art. For instance,in particular embodiments, the biofilm disruptor composition may becontained in the tuft holes of the toothbrush. Alternatively, in otherembodiments, the biofilm disruptor composition may be coated or embeddedin the bristles of the toothbrush. Optionally, other parts of thetoothbrush may also be coated or embedded with the biofilm disruptorcomposition, including any parts of the toothbrush that supplement thebristles and are designed to be contacted with the oral cavity. Incertain embodiments, toothbrushes may contain rubber paddles, tonguecleaners, or other pieces extended from the head for the purposes ofbeing contacted with the tooth, tongue, gums, or other areas of the oralcavity. In such embodiments, these parts may be embedded with thebiofilm disruptor composition and, optionally, a surfactant, biocide,and/or other additive.

In certain embodiments, the biofilm disruptor may also be incorporatedinto or used to form an encapsulated system to allow for a controlledrelease. In these embodiments, the biofilm disruptor composition canoptionally be in the form of a plurality of small microspheres thatencapsulate the biofilm disruptor. The microspheres can optionally havean outer coating of dissolvable material that enables the biofilmdisruptor to slowly release over repeated brushings.

In certain embodiments, the present invention also relates to a methodof cleaning and/or disinfecting articles such as contact lenses. Themethod of these embodiments involves treating contact lenses with acleaning and/or disinfecting solution containing the biofilm disruptoraccording to the present invention. In some embodiments, the contactlens may be treated in this manner while being stored in solution orwhile being used in vivo. In alternative embodiments, the biofilmdisruptor can be used in eye drops.

In certain embodiments, the present invention further relates to amethod of treating and/or preventing acne or other biofilm-associatedskin infections on the skin of a subject. The methods of theseembodiments involve treating the skin of the subject systemically or theskin surface topically with the biofilm disruptor according to thepresent invention under conditions effective to treat and/or prevent theacne or biofilm-associated skin infections. In some embodiments, thebiofilm disruptor may be present in an ointment, cream, liniment,salves, shaving lotion, or aftershave. In these embodiments, the biofilmdisruptor may also be present in a powder, cosmetic, ointment, cream,liquid, soap, gel, suspension, lotion, solution, paste, spray, aerosol,oil, cosmetic applicator, and/or solid, woven or non-woven materialintended to contact or be proximate with the skin. In other embodiments,biofilm disruptor may be present in suspensions, syrups, elixirs,solutions, pills, capsules, suppositories and tablets for oral systemicuse.

In certain embodiments, the present invention also relates to a methodof treating and/or preventing a chronic biofilm-associated disease in asubject. The methods of these embodiments involve administering to thesubject the biofilm disruptor according to the present invention underconditions effective to treat and/or prevent the chronicbiofilm-associated disease. The chronic biofilm-associated diseases tobe treated and/or prevented include, but are not limited to, middle earinfections, osteomyelitis, prostatitis, colitis, vaginitis, urethritis,arterial plaques, sinovial infections, infections along tissue fascia,respiratory tract infections (e.g., infections associated with lunginfections of cystic fibrosis patients, pneumonia, pleurisy, pericardialinfections), genito-urinary infections, and gastric or duodenal ulcerinfections. In some embodiments, the biofilm disruptor may beadministered in combination with the antimicrobial actives describedabove. In one embodiment, the biofilm disruptor and the antimicrobialtreatment are administered simultaneously. In another embodiment, thebiofilm disruptor and antimicrobial treatment are administeredseparately. In the case of gastric therapies, gastrointestinal activesmay also be employed as described in U.S. Pat. No. 6,986,901 to Meiselet al., herein incorporated by reference in its entirety.

The biofilm disruptor can be impregnated in a surface in order toinhibit formation of a biofilm on the surface. Alternatively, thebiofilm disruptor can be in a copolymer or a gel coating over thesurface.

In certain embodiments, the present invention further relates to kitscomprising the biofilm disruptor and instructions on the use of thebiofilm disruptor with the packages containing the biofilm disruptor orwith other forms of advertising associated with the sale or use of thebiofilm disruptor. In certain embodiments, the instructions may beincluded in any manner typically used by consumer product manufacturingor supply companies. Nonlimiting examples include providing instructionson a label attached to the container holding the compounds and/orcompositions; on a sheet either attached to the container oraccompanying it when purchased; or in advertisements, demonstrations,and/or other written or oral instructions which may be connected to thepurchase or use of the compounds and/or compositions.

In particular embodiments, the instructions will include a descriptionof the use of the biofilm disruptors. In these embodiments, theinstructions, for instance, may additionally include informationrelating to the recommended amount of compounds and/or compositions toapply to the substrate.

The compounds and compositions described herein can be prepared byconventional organic syntheses, readily available to one of ordinaryskill in the art without undue experimentation. Specific examples aredescribed herein below.

EXAMPLES Example 1

Screening of Biofilm Disruptive Agents

Formulations containing unsaturated long chain alcohols as the biofilmdisruptive agents were prepared according to the following formula:

Raw material % (w/v) Water 77.75 Ethanol (190 proof) 22 Unsaturated longchain alcohol 0.05 Pluronic F-127 0.2

Separate formulations were prepared from the above ingredients with eachformulation containing one of the unsaturated long chain alcohols inTable 1. The formulations were then tested as indicated below. Inaddition, the following negative control without any unsaturated longchain alcohol was also prepared.

Raw material % (w/v) Water 77.80 Ethanol (190 proof) 22 Pluronic F-1270.2

A 48-hour salivary biofilm was grown on a polystyrene peg platesubstrate (96 pegs, N=8 per test group). The pegs were subsequentlytreated for twenty minutes with each of the long chain alcoholsformulations described above, as well as the negative control. Aftertreatment, viable bacteria remaining on the substrate were removed bysonication using a Ultrasonic processor XL by Misonix (Farmingdale,N.Y.), lysed with Celsis Luminex (Celsis Rapid Detection Rapiscreen(Celsis International PLC, Chicago)) and ATP from the bacteria weremeasured using a bioluminescence marker Celsis LuminATE (Celsis RapidDetection Rapiscreen (Celsis International PLC, Chicago)). Data wasreported in log RLU (relative light units), where decreasing log RLUsindicates fewer bacteria remaining on the substrate and in M-factorunits where M-factor is the difference between the log RLU values of thecompound tested and the negative control. The results of the test areshown on Table 1.

TABLE 1 Biofilm treatment test results log RLU Unsaturated Long Supplierlog RLU negative M- Chain Alcohol Information compound control factorcis-2-hexen-1-ol Sigma-Aldrich 5.21 5.32 0.11 St. Louis, MO 63103cis-3-hexen-1-ol TCI America 5.22 5.28 0.06 Portland, OR 97203trans-2-hexen-1-ol SAFC (Sigma 5.20 5.32 0.12 Aldrich) St. Louis, MO63103 cis-3-hepten-1-ol SAFC (Sigma 5.07 5.32 0.25 Aldrich) St. Louis,MO 63103 cis-5-octen-1-ol, SAFC (Sigma 5.01 5.32 0.31 Aldrich) St.Louis, MO 63103 trans-2-octen-1-ol Sigma-Aldrich 4.89 5.58 0.69 St.Louis, MO 63103 Alfa Aesar Ward Hill, MA 01835 1-octen-3-ol AcrosOrganics 4.99 5.32 0.33 US Distributor: Morris Plains, NJ 07950cis-2-nonen-1-ol SAFC (Sigma 4.57 5.33 0.76 Aldrich) St. Louis, MO 63103cis-3-nonen-1-ol Sigma-Aldrich 4.56 5.32 0.76 St. Louis, MO 63103cis-6-nonen-1-ol SAFC (Sigma 4.60 5.32 0.72 Aldrich) St. Louis, MO 63103trans-2-nonen-1-ol SAFC (Sigma 4.83 5.28 0.45 Aldrich) St. Louis, MO63103 cis-4-decen-1-ol Sigma-Aldrich 4.71 5.33 0.62 St. Louis, MO 63103trans-2-decen-1-ol Sigma-Aldrich 4.13 5.58 1.45 St. Louis, MO 631039-decen-1-ol Sigma-Aldrich 4.35 5.32 0.97 St. Louis, MO 631031-decen-3-ol SAFC (Sigma 4.46 5.32 0.86 Aldrich) St. Louis, MO 63103trans-2-dodecen-1-ol Fluka (Sigma- 4.44 5.32 0.88 Aldrich) St. Louis, MO63103 trans-2-undecen-1-ol Alfa Aesar 4.72 5.28 0.56 Ward Hill, MA 01835

Table 1 shows that unsaturated alcohols of chain length of at least 7carbons resulted in an activity difference versus the negative control(M-factor) of greater than 0.21.

Example 2

Screening of Biofilm Disruptive Agents

Formulations containing long chain aldehydes as the biofilm disruptiveagents were prepared according to the following formula:

Raw material % (w/v) Water 77.75 Ethanol (190 proof) 22 Unsaturated longchain aldehyde 0.05 Pluronic F-127 0.2

Separate formulations were prepared from the above ingredients with eachformulation containing one of the unsaturated long chain aldehydes inTable 2. The formulations were then tested as indicated below. Inaddition, the following negative control without any unsaturated longchain aldehyde was also prepared.

Raw material % (w/v) Water 77.80 Ethanol (190 proof) 22 Pluronic F-1270.2

A 48-hour salivary biofilm was grown on a polystyrene peg platesubstrate (96 pegs, N=8 per test group). The pegs were subsequentlytreated for twenty minutes with each of the long chain alcoholsformulations described above, as well as the negative control. Aftertreatment, viable bacteria remaining on the substrate were removed bysonication using a Ultrasonic processor XL by Misonix (Farmingdale,N.Y.), lysed with Celsis Luminex (Celsis Rapid Detection Rapiscreen(Celsis International PLC, Chicago)) and ATP from the bacteria weremeasured using a bioluminescence marker. Celsis LuminATE (Celsis RapidDetection Rapiscreen (Celsis International PLC, Chicago)). Data wasreported in log RLU (relative light units), where decreasing log RLUsindicates fewer bacteria remaining on the substrate and in M-factorunits where M-factor is the difference between the log RLU values of thecompound tested and the negative control. The results of the test areshown on Table 2.

TABLE 2 Biofilm Treatment Test Results log RLU Unsaturated Long Supplierlog RLU negative M- Chain Aldehyde Information compound control factortrans-2-hexenal SAFC (Sigma 5.39 5.58 0.19 Aldrich) St. Louis, MO 63103trans-2-heptenal SAFC (Sigma 5.52 5.58 0.06 Aldrich) St. Louis, MO 63103cis-4-hepten-1-al TCI America 5.14 5.10 −0.04 Portland, OR 972032,6-dimethyl-5- TCI America 5.07 5.28 0.21 hepten-1-al Portland, OR97203 trans-2-octenal SAFC (Sigma 5.26 5.58 0.32 Aldrich) St. Louis, MO63103 trans-2-nonenal SAFC (Sigma 4.66 5.58 0.92 Aldrich) St. Louis, MO63103 cis-6-nonenal Alfa Aesar 4.84 5.28 0.44 Ward Hill, MA 01835cis-4-decenal SAFC (Sigma 4.59 5.58 0.99 Aldrich) St. Louis, MO 63103cis-7-decenal Sigma Aldrich 5.25 5.58 0.33 St. Louis, MO 63103trans-2-decenal Fluka (Sigma 4.63 5.58 0.95 Aldrich) St. Louis, MO 63103trans-2-undecenal SAFC (Sigma 4.49 5.32 0.83 Aldrich) St. Louis, MO63103 trans-2-dodecenal SAFC (Sigma 4.58 5.58 1.00 Aldrich) St. Louis,MO 63103

Table 2 shows that unsaturated aldehydes of chain length of at least 7carbons resulted in an activity difference versus the negative control(M-factor) of greater than 0.21.

Example 3

Concentration Dependence of Biofilm Disruptor Activity

To determine the concentration dependence of biofilm disruptor activity,a series of formulations was prepared with cis-2-nonen-1-olconcentration ranging from 0.0005% to 0.1%. These formulations weretested against the negative control in the biofilm disruption screeningassay described in Examples 1 and 2. Data was reported in log RLU(relative light units) where decreasing log RLUs indicates fewerbacteria remaining on the substrate and in M-factor units where M-factoris the difference between the log RLU values of the compound tested andthe negative control. The results of the test are shown on Table 3.

TABLE 3 Biofilm treatment results as a function of concentration forcis-2-nonen-1-ol. Negative A % B % C % D % E % F % G % control Rawmaterial (w/v) (w/v) (w/v) (w/v) (w/v) (w/v) (w/v) % (w/v) Water 77.777.75 77.775 77.79 77.795 77.799 77.7995 77.8 Ethanol (190 22 22 22 2222 22 22 22 proof) cis-2-nonen-1- 0.1 0.05 0.025 0.01 0.005 0.001 0.00050 ol Pluronic F-127 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 log RLU 4.81 4.825.28 5.59 5.69 5.70 5.72 5.63 M-factor 0.82 0.82 0.35 0.04 −0.06 −0.07−0.09 NA

These results indicate that biofilm disruption improves (i.e., M-factorincreases) with increasing concentration of the biofilm disruptor.

Example 4

Combinations of Long-Chain Alcohols

To further evaluate the potential of combining two biofilm disruptiveagents, the following formulations containing cis-2-nonen-1-ol andcis-6-nonen-1-ol were prepared. These formulations were tested againstthe negative control in the biofilm disruption screening assay describedin Examples 1 and 2. Data was reported in log RLU (relative light units)where log RLUs indicates the amount of bacteria remaining on thesubstrate. The results of the test are shown on Table 4.

TABLE 4 Biofilm treatment results in blends of cis-2-nonen-1-ol andcis-6-nonen-1-ol. A % B % C % D % E % Raw material (w/v) (w/v) (w/v)(w/v) (w/v) Water 77.75 77.75 77.75 77.75 77.75 Ethanol (190 proof) 2222 22 22 22 cis-2-nonen-1-ol 0.05 0.01 0 0.04 0.025 cis-6-nonen-1-ol 00.04 0.05 0.01 0.025 Pluronic F-127 0.2 0.2 0.2 0.2 0.2 log RLU 4.754.86 4.78 4.9 4.81

Table 4 indicates effectiveness (i.e., similarly reduced log RLU values)of using either individual biofilm disruptors or combinations of thebiofilm disruptors.

Example 5

Combinations of Biofilm Disruptors and Antimicrobial Actives

The following formulations containing antimicrobial actives (menthol,thymol, eucalyptol, and methyl salicylate) were prepared and evaluatedin a flow-through biofilm model to evaluate potential for oral plaquedisruption following a thirty-second treatment.

Wildtype organisms harvested from human saliva inoculate media flowingpast pegs on a 96-peg polystyrene plate where organisms adhere anddevelop into biofilms. The biofilms are treated twice a day for a totalof five treatments over the course of 60 hours.

Data was reported in log RLU (relative light units) where decreasing logRLUs indicates fewer bacteria remaining on the substrate. The results ofthe test are shown on Table 4.

TABLE 5 Biofilm treatment results of blends of cis-2-nonen-1-ol and1-decen-3-ol in formulations containing anti-microbial agents (menthol,thymol, eucalyptol, and methyl salicylate). Raw material A (% w/v) B (%w/v) C (% w/v) L-Menthol, USP 0.042 0.042 0.042 Thymol NF 0.064 0.0640.064 Methyl Salicylate NF 0.066 0.066 0.066 Eucalyptol USP 0.092 0.0920.092 Pluronic F-127 NF Prill 0.250 0.250 0.250 Benzoic Acid 0.128 0.1200.120 Sodium Benzoate 0.026 0.0354 0.0354 Sodium saccharin 0.020 0.1170.117 Sucralose 0.041 0 0 FD&C Green No. 3 0.0005 0.0005 0.0005 Sorbitol70%, USP 20.0 20.0 20.0 Alcohol, 190 proof 21.6 (% v/v) 21.6 (% v/v)21.6 (% v/v) Flavor 0.12 0.085 0.085 cis-2-nonen-1-ol 0.05 0 01-decen-3-ol 0 0.05 0 Purified Water 79.1 79.1 79.1 TOTAL 100 100 100log RLU 4.46 4.36 4.75

Formulations A and B, containing biofilm disrupting agentscis-2-nonen-1-ol and 1-decen-3-ol respectively, results in reducedbiofilm remaining on the substrate (i.e., decreased log RLU) whencompared to the Formulation C control (log RLU=4.75).

Various embodiments of the invention have been set forth above. Eachembodiment is provided by way of explanation of the invention, notlimitation of the invention. In fact, it will be apparent to thoseskilled in the art that various modifications and variations can be madein the present invention without departing from the scope or spirit ofthe invention. For instance, features illustrated or described as partof one embodiment, can be used on another embodiment to yield a stillfurther embodiment. Thus, it is intended that the present inventioncover such modifications and variations as come within the scope of theappended claims and their equivalents.

What is claimed is:
 1. A method of disrupting biofilms, comprising thesteps of: i. providing a surface or substrate comprising a biofilm; andii. applying to the surface or substrate from about 0.005% to about 10%of a biofilm disruptor comprising a compound selected from the groupconsisting of 1-decen-3-ol; cis-2-nonen-1-ol, and mixtures thereof. 2.The method according to claim 1, wherein the biofilm disruptor is1-decen-3-ol.
 3. The method according to claim 1, wherein the surface isany surface in the oral cavity and the biofilm disruptor is present at aconcentration of 0.005% to 0.5%.
 4. The method according to claim 1,wherein the surface is a mammalian or human skin or mucosal surface andthe biofilm disruptor is present at a concentration of 0.005% to 5%. 5.The method of claim 1 wherein the biofilm disruptor comprisescis-2-nonen-1-ol.